title: Alteration in cellular viability, pro-inflammatory cytokines and nitric oxide production in nephrotoxicity generation by Amphotericin B: involvement of PKA pathway signaling.
abstract: Amphotericin B is one of the most effective antifungal agents; however, its use is often limited owing to adverse effects, especially nephrotoxicity. The purpose of this study was to evaluate the effect of inhibiting the PKA signaling pathway in nephrotoxicity using Amphotericin B from the assessment of cell viability, pro-inflammatory cytokines and nitric oxide (NO) production in LLC-PK1 and MDCK cell lines. Amphotericin B proved to be cytotoxic for both cell lines, as assessed by the mitochondrial enzyme activity (MTT) assay; caused DNA fragmentation, determined by flow cytometry using the propidium iodide (PI) dye; and activated the PKA pathway (western blot assay). In MDCK cells, the inhibition of the PKA signaling pathway (using the H89 inhibitor) caused a significant reduction in DNA fragmentation. In both cells lines the production of interleukin-6 (IL)-6 proved to be a dependent PKA pathway, whereas tumor necrosis factor-alpha (TNF-α) was not influenced by the inhibition of the PKA pathway. The NO production was increased when cells were pre-incubated with H89 followed by Amphotericin B, and this production produced a dependent PKA pathway in LLC-PK1 and MDCK cells lines. Therefore, considering the present study's results as a whole, it can be concluded that the inhibition of the PKA signaling pathway can aid in reducing the degree of nephrotoxicity caused by Amphotericin B.
authors.affiliation: Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antônio Carlos 6627, 30161-970, Belo Horizonte, MG, Brasil.
authors.forename: F D
authors.initials: FD
authors.lastname: França
authors.forename: A F
authors.initials: AF
authors.lastname: Ferreira
authors.forename: R C
authors.initials: RC
authors.lastname: Lara
authors.forename: J V
authors.initials: JV
authors.lastname: Rossoni
authors.forename: D C
authors.initials: DC
authors.lastname: Costa
authors.forename: K C M
authors.initials: KC
authors.lastname: Moraes
authors.forename: C A
authors.initials: CA
authors.lastname: Tagliati
authors.forename: M M
authors.initials: MM
authors.lastname: Chaves
mesh_terms: D000666:Amphotericin B / Q000633:toxicity*; D000818:Animals; D000935:Antifungal Agents / Q000633:toxicity*; D018929:Cell Culture Techniques; D002470:Cell Survival / Q000187:drug effects; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D016207:Cytokines / Q000096:biosynthesis*; D053938:DNA Fragmentation / Q000187:drug effects; D004285:Dogs; D015850:Interleukin-6 / Q000096:biosynthesis; D007668:Kidney / Q000187:drug effects* / Q000201:enzymology / Q000276:immunology / Q000473:pathology; D018374:LLC-PK1 Cells; D061985:Madin Darby Canine Kidney Cells; D009569:Nitric Oxide / Q000096:biosynthesis*; D015398:Signal Transduction; D013552:Swine; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
chemical_list: D000935:Antifungal Agents; D016207:Cytokines; D015850:Interleukin-6; D014409:Tumor Necrosis Factor-alpha; D009569:Nitric Oxide; D000666:Amphotericin B; D017868:Cyclic AMP-Dependent Protein Kinases
keywords: IL-6; LLC-PK1; MDCK; NO; PKA; TNF-α; amphotericin B; nephrotoxicity